苹果砧木SH40 MdNCED1基因克隆与表达分析
作者:
作者单位:

作者简介:

通讯作者:

中图分类号:

基金项目:

国家科技攻关计划


Isolation and Expression of MdNCED1 Gene from Apple Rootstock SH40
Author:
Affiliation:

Fund Project:

The National Key Technologies R&D Program of China

  • 摘要
  • |
  • 图/表
  • |
  • 访问统计
  • |
  • 参考文献
  • |
  • 相似文献
  • |
  • 引证文献
  • |
  • 资源附件
  • |
  • 文章评论
    摘要:

    以苹果砧木SH40(Malus domestica ?Malus Honanensis)为试材,采用RT-PCR结合RACE技术从苹果茎尖中克隆得到一个脱落酸(ABA)代谢合成关键基因:9-顺式-环氧类胡萝卜素双加氧酶(NCED)基因cDNA序列全长,命名为MdNCED1,其全长2179bp,编码606个氨基酸。该基因在GenBank登录号为KC816734。氨基酸同源性分析表明,MdNCED1与已报道的其他植物物种的氨基酸序列具有63.7%-93.0%的相似性。通过构建MdNCED1基因的原核表达载体pDEST15-MdNCED1,转入大肠杆菌(DE3),用IPTG诱导。SDS-PAGE分析表明,MdNCED1基因在大肠杆菌中被诱导表达蛋白质分子量与预期结果一致。荧光定量结果表明,MdNCED1基因在SH28、M26、SH40及其嫁接品种“嘎啦”的表达趋势均呈先上升后下降的过程,同时与其矮生程度呈正相关。

    Abstract:

    9-cis-epoxycarotenoid (NCED) gene is a critical gene in the pathway of abscisic acid biosynthesis. In this research, NCED gene in apple, designated as MdNCED1 (GenBank accession number: KC816734), was isolated from the apical tissue of apple rootstock SH40 (M. domestica ?M. Honanensis)by RT-PCR and RACE technology. The full cDNA was 2179bp,containing a complete open reading frame that encodes 606 amino acids. Amino acid sequence analysis revealed that the sequence had 63.7%-93.0% similarity with those of other reported plants. The ORF fragment of the gene was cloned into vector pDEST15,the recombined plasmid was transformed in to E. coli. (DE3). The result of SDS-PAGE demonstrated that the size of expected protein in the prokaryotic expression system. Fluorescent quantitative PCR analysis showed that the trend of expression of SH28, M26, SH40 and the grafted varieties ‘Gala’ were increased and then decreased. And the expression of this gene was positively associated to the dwarfing degree.

    参考文献
    相似文献
    引证文献
引用本文

姜志昂,孙建设,彭建营,等.苹果砧木SH40 MdNCED1基因克隆与表达分析[J].植物遗传资源学报,2014,15(1):154-160.

复制
分享
文章指标
  • 点击次数:
  • 下载次数:
  • HTML阅读次数:
  • 引用次数:
历史
  • 收稿日期:2013-04-26
  • 最后修改日期:2013-06-24
  • 录用日期:2013-12-10
  • 在线发布日期: 2013-12-26
  • 出版日期:
文章二维码
您是第位访问者
ICP:京ICP备09069690号-23
京ICP备09069690号-23
植物遗传资源学报 ® 2024 版权所有
技术支持:北京勤云科技发展有限公司